Towards sustainable and humane dairy farming: A low-cost electrochemical sensor for on-site diagnosis of milk fever.

Milk fever is a metabolic disorder that predominantly affects dairy animals during the periparturient period and within four weeks of calving. Milk fever is primarily attributed to a decrease in the animal's serum Ca2+ levels. Clinical milk fever occurs when Ca2+ concentration drops below 1.5 mM (6 mg/dL). Without prompt intervention, clinical milk fever leads to noticeable physical symptoms and health complications including coma and fatality. Subclinical milk fever is characterized by Ca2+ levels between 1.5 and 2.12 mM (6-8.48 mg/dL). Approximately 50% of multiparous dairy cows suffer from subclinical milk fever during the transition to lactation. The economic impact of milk fever, both direct and indirect, is substantial, posing challenges for farmers. To address this issue, we developed a low-cost electrochemical sensor that can measure bovine serum calcium levels on-site, providing an opportunity for early detection of subclinical and clinical milk fever and early intervention. This calcium sensor is a scalable solid contact ion sensing platform that incorporates a polymeric calcium-selective membrane and ionic liquid-based reference membrane into laser-induced graphene (LIG) electrodes. Our sensing platform demonstrates a sensitivity close to the theoretical Nernstian value (29.6 mV/dec) with a limit of detection of 15.6 µM and selectivity against the species in bovine serum. Moreover, our sensor can detect Ca2+ in bovine serum with 91% recovery.

SPOOC (Sensor for Periodic Observation of Choline): An Integrated Lab-on-a-Spoon Platform for At-Home Quantification of Choline in Infant Formula.

Choline is an essential micronutrient for infants' brain development and health. To ensure that infants receive the needed daily dose of choline, the U.S. Food and Drug Administration (FDA) has set requirements for choline levels in commercialized infant formulas. Unfortunately, not all families can access well-regulated formulas, leading to potential inadequacies in choline intake. Economic constraints or difficulties in obtaining formulas, exacerbated by situations like COVID-19, prompt families to stretch formulas. Accurate measurement of choline in infant formulas becomes imperative to guarantee that infants receive the necessary nutritional support. Yet, accessible tools for this purpose are lacking. An innovative integrated sensor for the periodic observation of choline (SPOOC) designed for at-home quantification of choline in infants' formulas and milk powders is reported. This system is composed of a choline potentiometric sensor and ionic-liquid reference electrode developed on laser-induced graphene (LIG) and integrated into a spoon-like device. SPOOC includes a micro-potentiometer that conducts the measurements and transmits results wirelessly to parents' mobile devices. SPOOC demonstrated rapid and accurate assessment of choline levels directly in pre-consuming infant formulas without any sample treatment. This work empowers parents with a user-friendly tool for choline monitoring promoting informed nutritional decision-making in the care of infants.

LiFT (a Lithium Fiber-Based Test): An At-Home Companion Diagnostics for a Safer Lithium Therapy in Bipolar Disorder.

This work presents LiFT (a lithium fiber-based test), a low-cost electrochemical sensor that can measure lithium in human saliva and urine with FDA-required accuracy. Lithium is used for the treatment of bipolar disorder, and has a narrow therapeutic window. Close monitoring of lithium concentration in biofluids and adjustment of drug dosage can minimize the devastating side effects. LiFT is an inexpensive, yet accurate and simple-to-operate lithium sensor for frequent at-home testing for early identification of lithium toxicity. The low cost and high accuracy of LiFT are enabled through an innovative design and the use of ubiquitous materials such as yarn and carbon black for fabrication. LiFT measures Li+ through potentiometric recognition using a lithium selective sensing membrane that is deposited on the ink-coated yarn. A detection limit of 0.97 µM is obtained with a sensitivity of 59.07±1.25 mV/decade for the Li+ sensor in deionized water. Moreover, the sodium correction extended LiFT's linear range in urine and saliva to 0.5 mM. The LiFT platform sends the test results to the patient's smartphone, which subsequently can be shared with the patient's healthcare provider to expedite diagnosis and prevention of acute lithium toxicity.

NF-kB affects migration of vascular smooth muscle cells after treatment with heparin and ibrutinib.

The migration of vascular smooth muscle cells (VSMCs) is one of the most important events in the remodeling of atherosclerosis plaque. The aim of study was to investigate the role of Heparin in the VSMC migration and its association with the NF-kB, collagen 1 and collagen 3 expression levels. Moreover, the incorporation of Heparin was studied in the VSMC cultures including Betulinic acid and Ibrutinib. Twelve cell groups were cultured and treated with the Heparin, Betulinic acid and Ibrutinib based on the viability and toxicity in 24-h and 48-h periods. The gene and protein expression levels were measured by RT-qPCR and western blotting techniques. The VSMC migration was determined by scratch test. In contrast with Ibrutinib (2 µM), Heparin (30 IU) increased significantly (P < 0.05) the NF-kB gene and protein expression levels and the VSMC migration during the exposure periods. Heparin (15 IU and 30 IU) also increased the collagen 1 gene expression level in the 48-h period while Heparin (5 IU and 15 IU) increased the collagen 3 gene expression levels in both periods. Incorporating Heparin into the cultures including Betulinic acid and Ibrutinib affected the collagen 1 and collagen 3 expression levels. The data suggested that the cell migration relates to NF-kB in the VSMCs treated with Heparin and Ibrutinib. Furthermore, the Heparin doses (5 IU and 15 IU) were safe for VSMCs based on the NF-kB, and collagen 3 expression levels.

The contribution of childhood maltreatment to the incidence of heavy cannabis use in Iran (IRNS-CCI): A multicenter, matched-pairs, nested, case-control study.

BACKGROUND: Previous studies have shown the role of the interaction between the endocannabinoid system (ECS) and life's adversities in the formation of addiction, including alcohol abuse. OBJECTIVE: Our objective was to identify childhood maltreatment (CM) patterns with the strongest impact on the probability of heavy cannabis use (THCCOOH concentrations ≥150 ng/mL) in Iran. PARTICIPANTS AND SETTING: Using survivor sampling, 350 adult participants were selected, and they were then allocated to three categories based on an optimal algorithm: 1) Sexual abuse, 2) Physical abuse, and 3) Physical neglect. METHODS: From 1 September 2019 to 1 May 2023, we implemented a multicenter, matched-pairs, nested, case-control study based on the wave 3-wave 6 data of a longitudinal, multicenter, cohort study. The cases and controls (n = 350 men) were defined according to the severity of CM. The THC potency was evaluated with the delta-9 carboxy tetrahydrocannabinol (THC-COOH) levels in urine using gas chromatography/mass spectrometry (GC/MS). We calculated the population attributable fractions (PAFs) to identify the patterns of maltreatment associated with the highest odds of high-potency cannabis use. RESULTS: Accumulating CM, including sexual abuse, physical abuse, and physical neglect, carried more than three times the risk of heavy cannabis use (OR 3.4 95 % CI 2.9-4.1), and the combination of the three indicators of maltreatment and a high BMI (25-29.9) carried more than four times the risk of heavy cannabis use (OR 4.7 95 % CI 2.7-4.1) compared to the controls. We estimated that in the case of zero CM for each of the three indicators, over 20 % of new cases of heavy cannabis use can be prevented. CONCLUSIONS: The findings show the significance of CM as a predicator of heavy cannabis use in adulthood and in the abstinence phase.

Beta arrestin-related signalling axes are influenced by dexamethasone and metformin in vascular smooth muscle cells cultured in high glucose condition.

BACKGROUND: Metformin (Met) and dexamethasone (Dexa) are known to reduce blood sugar levels and anti-inflammatory effects, respectively. Based on the acceleration of atherosclerosis process in diabetes, the ß-arrestin 2 (BARR2) gene and protein expression levels were evaluated in vascular smooth muscle cells (VSMCs) treated with Met and Dexa in high glucose conditions in this study. METHODS AND MATERIALS: Human VSMCs were cultured in Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM-F12) medium and, were treated with different values of Met (1 mM, 5 mM and 7 mM) and Dexa (10-7 M, 10-6 M and 10-5 M) in 24- and 48-h periods. The BARR2 gene and protein expression levels were identified with RT-qPCR and western blotting techniques, respectively. The signalling axes were predicted from gene network made using Cytoscape software and, were annotated with Gene Ontology. RESULTS: The BARR2 gene and protein expression levels reduced in VSMCs treated with Dexa and Met after 24- and 48-h periods. These results were more changed after 48 h. Furthermore, many BARR2-related signalling axes were found from the network genes. CONCLUSION: Met and Dexa suppressed the BARR2 protein and gene expression levels in the VSMCs. Moreover, the gene network suggested some the cellular signalling axes related to BARR2 that may be affected by Met and Dexa.

Recent Progress on Green New Phase Extraction and Preparation of Polyphenols in Edible Oil.

With the proposal of replacing toxic solvents with non-toxic solvents in the concept of green chemistry, the development and utilization of new green extraction techniques have become a research hotspot. Phenolic compounds in edible oils have good antioxidant activity, but due to their low content and complex matrix, it is difficult to achieve a high extraction rate in a green and efficient way. This paper reviews the current research status of novel extraction materials in solid-phase extraction, including carbon nanotubes, graphene and metal-organic frameworks, as well as the application of green chemical materials in liquid-phase extraction, including deep eutectic solvents, ionic liquids, supercritical fluids and supramolecular solvents. The aim is to provide a more specific reference for realizing the green and efficient extraction of polyphenolic compounds from edible oils, as well as another possibility for the future research trend of green extraction technology.

FAST (Flexible Acetylcholine Sensing Thread): Real-Time Detection of Acetylcholine with a Flexible Solid-Contact Potentiometric Sensor.

Acetylcholine (ACh) is involved in memory and learning and has implications in neurodegenerative diseases; it is therefore important to study the dynamics of ACh in the brain. This work creates a flexible solid-contact potentiometric sensor for in vitro and in vivo recording of ACh in the brain and tissue homogenate. We fabricate this sensor using a 250 µm diameter cotton yarn coated with a flexible conductive ink and an ACh sensing membrane that contains a calix[4]arene ionophore. The exposed ion-to-electron transducer was sealed with a 2.5 µm thick Parylene C coating to maintain the flexibility of the sensor. The resulting diameter of the flexible ACh sensing thread (FAST) was 400 µm. The FAST showed a linear response range from 1.0 µM to 10.0 mM in deionized water, with a near-Nernstian slope of 56.11 mV/decade and a limit of detection of 2.6 µM. In artificial cerebrospinal fluid, the limit of detection increased to 20 µM due to the background signal of ionic content of the cerebrospinal fluid. The FAST showed a signal stability of 226 µV/h over 24 h. We show that FAST can measure ACh dynamics in sheep brain tissue and sheep brain homogenate after ACh spiking. FAST is the first flexible electrochemical sensor for monitoring ACh dynamics in the brain.

The Potential Relationship Between Serum Irisin Concentration With Inflammatory Cytokines, Oxidative Stress Biomarkers, Glycemic Indices and Lipid Profiles in Obese Patients With Type 2 Diabetes Mellitus: A Pilot Study.

Objectives: Diabetes mellitus is a serious health-treated problem identified by disorders such as insulin resistance, dyslipidemia, and inflammation. Irisin, a newly discovered myokine/adipokine, is involved in metabolic homeostasis. The present study was carried out to investigate the potential relationship between serum irisin with inflammatory cytokines, oxidative stress biomarkers, glycemic indices, and lipid profiles in obese patients with type 2 diabetes mellitus. Methodology: This analytical cross-sectional study was conducted on 62 participants (n=32 obese participants with diabetes, n=30 participants with normal weight). The participants answered a demographic questionnaire. Serum irisin, glycemic indices, lipid profiles, inflammatory cytokines and oxidative stress biomarkers were measured using standard methods. The difference between groups was assessed by independent-sample t-test or by a non-parametric equivalent. For qualitative variables, the Chi-Square test was used. Pearson rho coefficient was used to determine the potential relationship between irisin and inflammatory cytokines, oxidative stress biomarkers, glycemic indices, and lipid profiles. A p<0.05 was defined as significant. Results: The median (IQR) age of the obese participants with diabetes was 54.0 years (52.2-60.7) and in the normal weight group was 38.0 years (30.0-47.2) (p<0.001). About 78% and 60% of participants in the obese with diabetes and the normal weight groups were females (p>0.05), respectively. Significant differences were observed in serum irisin levels between the two groups, with lower levels (218.74 ng/mL, [144.98-269.26]) noted in the obese with diabetes group compared to the normal weight group (266.68 ng/mL, [200.64-336.57]) with a p=0.024. There was a substantial difference between the two groups regarding IL-6, TNF-α, and hs-CRP (p<0.05). IL-6 had a moderate negative correlation with irisin in obese patients with T2DM (r=-0.478, p=0.006). Conclusion: Irisin concentration was detected to be lower in obese people with diabetes. A negative relationship was detected between irisin and IL-6. Considering emerging evidence about the beneficial functions of irisin in improving metabolic abnormalities, designing future studies with greater sample sizes that will validate these results is needed.

Impedimetric Sensing: An Emerging Tool for Combating the COVID-19 Pandemic.

The COVID-19 pandemic revealed a pressing need for the development of sensitive and low-cost point-of-care sensors for disease diagnosis. The current standard of care for COVID-19 is quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). This method is sensitive, but takes time, effort, and requires specialized equipment and reagents to be performed correctly. This make it unsuitable for widespread, rapid testing and causes poor individual and policy decision-making. Rapid antigen tests (RATs) are a widely used alternative that provide results quickly but have low sensitivity and are prone to false negatives, particularly in cases with lower viral burden. Electrochemical sensors have shown much promise in filling this technology gap, and impedance spectroscopy specifically has exciting potential in rapid screening of COVID-19. Due to the data-rich nature of impedance measurements performed at different frequencies, this method lends itself to machine-leaning (ML) algorithms for further data processing. This review summarizes the current state of impedance spectroscopy-based point-of-care sensors for the detection of the SARS-CoV-2 virus. This article also suggests future directions to address the technology's current limitations to move forward in this current pandemic and prepare for future outbreaks.

The effect of metformin on the metabolism of human vascular smooth muscle cells in high glucose conditions.

OBJECTIVES: Metformin is widely used in type 2 diabetic patients as an antihyperglycemic drug. The aim of this study was to investigate the effect of metformin on the metabolism of vascular smooth muscle cells in high glucose conditions. MATERIALS AND METHODS: The vascular smooth muscle cells were cultured in DMEM F12 containing glucose as high as 25 mM. The preconditioned cells were then treated with metformin in doses of 1, 5, and 7 mM for 24 h. MTT method was used to determine cell viability. Biochemical parameters including lactate, glucose, total protein, creatinine, and triglyceride were measured in the cell culture after the treatment with metformin. Oil Red O staining method was used to stain the lipids in the cells. RESULTS: Metformin reduced significantly (p<0.001) VSMC proliferation in a concentration-dependent manner. With the increase of glucose uptake by VSMCs, the cell lipid deposition was not changed. Other biochemical parameters such as lactate, triglyceride, total protein, and creatinine were significantly changed in the cell culture (p<0.05). CONCLUSIONS: Metformin increased the glucose uptake impacting metabolic pathways in VSMCs. It also increased the lactate efflux and protein metabolism without the change in cellular lipid deposition in high glucose conditions.

Non-coding RNAs are correlated to TGF-ß receptor type 2 in patients with colorectal cancer.

BACKGROUND: Multiple molecular expression alterations, particularly in non-coding RNAs, play fundamental roles in the regulations of cellular processes and may relate to the occurrence and progression of colorectal cancer (CRC). In the present study, we investigated the associations between TGFBR2, miR20a-5p and long non-coding RNA (lncRNA) LAMTOR5-AS1 in CRC patients. METHODS: Colorectal cancer and adjacent normal tissue samples (n = 34) were prepared from CRC patients. The associations between TGFBR2, miR20a-5p and lncRNA LAMTOR5-AS1 were predicted using bioinformatics tools. The expression levels of TGFBR2, miR20a-5p and lncRNA LAMTOR5-AS1 were measured using a quantitative real-time polymerase chain reaction technique. The TGFBR2 protein values were measured by western blotting. The clinical importance of lncRNA LAMTOR5-AS1 was assessed using receiver operating characteristic curve. RESULTS: The up-regulated levels of TGFBR2 (p = 0.02), TGFBR2 protein (p = 0.008) and lncRNA LAMTOR5-AS1 (p = 0.02) were significantly observed in CRC tissues compared to the adjacent normal tissues. The miR20a-5p expression level (p = 0.009) was downregulated in CRC tissues. In addition, the miR20a-5p expression level was inversely correlated to the TGFBR2 gene (r2  = 0.88, p < 0.0001), protein (r2  = 0.95, p < 0.0001) and lncRNA LAMTOR5-AS1 gene (r2  = 0.93, p < 0.0001) expression levels. Based on the area under curve, the increase of lncRNA LAMTOR5-AS1 expression level with a sensitivity of 64.52% and specificity of 65.52% was considered in CRC patients. CONCLUSIONS: We propose that miR20a-5p is inversely related to long non-coding RNA (lncRNA) LAMTOR5-AS, such that it may be involved in the regulation of TGFBR2 expression level in CRC patients.

Exploration of meteorin-like peptide (metrnl) predictors in type 2 diabetic patients: the potential role of irisin, and other biochemical parameters.

OBJECTIVES: Meteorin-like peptide (Metrnl), the newly discovered adipokines involves in glucose and lipid metabolism and energy homeostasis. The aim of the present study was to explore the potential predictors of Metrnl by emphasizing the Irisin, glycemic indices, and lipid profile biomarkers in type 2 diabetic patients. METHODS: This cross-sectional study was carried out on 32 obese types 2 diabetic patients, 31 healthy obese, and 30 healthy normal weight people between August 2020 and March 2021. Serum Metrnl and Irisin, fasting blood glucose (FBS), fasting insulin (FI), fasting insulin (FI), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), total cholesterol (TC), HbA1c and eAG levels were measured in a standard manner. To assay insulin resistance and insulin sensitivity, the homeostatic model assessment insulin resistance (HOMA-IR) and quantitative check index (QUICKI) model were used. Quantile regression analysis with the backward elimination method was used to explore predictors. The significant level was defined as p<0.05. RESULTS: Between variables entered into the model, only the group item showed to be the main predictor of Metrnl in type 2 diabetic patients. Besides, the serum level of Irisin was lower in diabetic patients, and a significant difference was detected between obese diabetic patients and the normal weight group (p=0.024). CONCLUSIONS: Given the multi-causality of diabetes and also the possible therapeutic role of Metrnl in the management of type 2 diabetic patients' abnormalities, designing future studies are needed to discover other predictors of Metrnl and the related mechanisms of Metrnl in the management of diabetes.

Comprehensive evaluation of the response to aluminum stress in olive tree (Olea europaea L.).

Olive (Olea europaea L.) is an ancient tree species in the Mediterranean, but the lack of knowledge about aluminum-resistant varieties limits its introduction to acidic soil. The objective of this study was to have a comprehensive evaluation of the response to aluminum stress in olive tree at germplasm, metabolome, and transcriptome levels. In this experiment, seedlings of 97 olive germplasm with 1.0-3.0 cm roots and two leaves were treated with 50 µM Al3+ (pH = 5.0). By factor analysis of the traits of defoliation rate, rooting rate, length of extended root, and length of new root, 97 germplasm were classified into five different groups according to their diverse responses to aluminum stress: 5 highly resistant (5.15%), 30 moderately resistant (30.93%), 31 general (31.96%), 23 moderately sensitive (23.71%), and 8 highly sensitive (8.25%) germplasm. The three most sensitive and three most resistant germplasm were further used for metabolome and transcriptome analysis. Exposed to aluminum stress, 96 differentially accumulated metabolites (DAMs)/4,845 differentially expressed genes (DEGs) and 66 DAMs/2,752 DEGs were identified in highly sensitive and resistant germplasm, respectively. Using multi-omics technology, the pathways and related DAMs/DEGs involved in cell wall/cytoplasm receptors, reactive oxygen species balance, hormone induction, synthesis of organic acids, Al3+ transport, and synthesis of metabolites were identified to mainly regulate the response to aluminum stress in olive. This study provides a theoretical guide and prior germplasm and genes for further genetic improvement of aluminum tolerance in the olive tree.

Dexamethasone suppresses the proliferation and migration of VSMCs by FAK in high glucose conditions.

BACKGROUND: High glucose conditions cause some changes in the vessels of diabetes through the signal transduction pathways. Dexamethasone and other corticosteroids have a wide range of biological effects in immunological events. In the present study, the effects of dexamethasone were investigated on the VSMC (vascular smooth muscle cell) proliferation, and migration based on the FAK gene and protein changes in high glucose conditions. METHODS AND MATERIALS: The vascular smooth muscle cells were cultured in DMEM and were treated with dexamethasone (10-7 M, 10-6 M, and 10-5 M) for 24, and 48 h in high glucose conditions. The cell viability was estimated by MTT method. The FAK gene expression levels and pFAK protein values were determined by RT-qPCR and western blotting techniques, respectively. A scratch assay was used to evaluate cellular migration. RESULTS: The FAK gene expression levels decreased significantly dependent on dexamethasone doses at 24 and 48 h. The pFAK protein values decreased significantly with a time lag at 24- and 48-h periods as compared with gene expression levels. CONCLUSION: The results showed that the inhibition of VSMC proliferation and migration by dexamethasone in the high glucose conditions may be related to the changes of FAK.

SARS-CoV-2 detection in hospital indoor environments, NW Iran.

This study aimed to investigate the potential contamination of SARS-CoV-2 in indoor settled dust and surfaces of Amir Al-Muminin hospital in Maragheh, Iran. Samples were taken from surfaces and settled dust using a passive approach and particulate matter (PM) using an active approach from different hospital wards. SARS-CoV-2 was detected in 15% of settled dust samples (N = 4/26) and 10% of surface samples (3/30). SARS-CoV-2 has been detected in 13.8% and 9.1% of the dust samples collected at a distance of fewer than 1 m and more than 3 m from the patient bed, respectively. SARS-CoV-2 was found in 11% of surface samples from low-touch surfaces and 8% from high touch surfaces. The relationship between PM2.5, PM10, humidity, temperature, and positive samples of SARS-CoV-2 was investigated. A positive correlation was observed between relative humidity, PM2.5, and positive SARS-CoV-2 samples. Principal component analysis (PCA) suggested positive correlation between positive SARS-CoV-2 samples, relative humidity, and PM2.5. Risk assessment results indicated that the annual mean infection risk of SARS-CoV-2 for hospital staff with illness and death was 2.6 × 10-2 and 7.7 × 10-4 per person per year. Current findings will help reduce the permanence of viral particles in the COVID 19 tragedy and future similar pandemics e.g., novel influenza viruses.

Characterization of plant growth-promoting rhizobacteria (PGPR) in Persian walnut associated with drought stress tolerance.

There is a lack of information on the rhizosphere of nut-bearing trees where microbial populations can benefit roots and tree growth. The current research aimed at discovering plant growth-promoting rhizobacteria (PGPR) in the rhizosphere of soil samples from around the root zone of six walnut trees, each of which was considered as a genotype, i.e. 'TT1', 'TT2', 'SS2', 'ZM1', 'Chandler' and 'Haward'. The trees grew in different arid and semiarid regions of Iran and Turkey. The strains were isolated and identified based on different morphological and biochemical markers. Drought-stress tolerance was assessed in the case of each isolate through their transfer to culture medium, containing polyethylene glycol (PEG6000) at 0 and 373.80 g L-1. Resilient strains were analyzed for measuring their ability to produce siderophore, hydrogen cyanide (HCN), Indole-3-acetic acid (IAA) and Gibberellic acid (GA3). In sum, 211 isolates were identified, of which a large number belonged to the Bacillus genus and, specifically, 78% of the strains were able to grow under drought stress conditions. The genus Arthrobacter was only detected in the rhizosphere of 'ZM1', 'Haward' and 'TT1' genotypes. In 4% of the strains, IAA production exceeded 53 mg L-1, while a high level of phosphorus solubility was verified in 6% of the strains. No strain was found to have the capability of producing HCN. The strains were screened for drought-tolerance, which resulted in the discovery of two promising strains, i.e. ZM39 and Cha43. Based on molecular identification through amplification and sequencing of the 16S rDNA gene, these two strains seemed to belong to Bacillus velezensis and Bacillus amyloliquefaciens, respectively. The discovery of new PGPR strains could probably assist walnut trees in improving their mechanisms of adaptation to drought stress.